Modulation of Enzyme Cascade Activity by Local Substrate Enrichment and Exclusion on DNA Nanostructures
Modulation of Enzyme Cascade Activity by Local Substrate Enrichment and Exclusion on DNA Nanostructures
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通过局部底物富集和排除 DNA 纳米结构来调节酶级联活性
DOI：
10.1021/acs.langmuir.2c02064
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发表时间：
2022
期刊：
影响因子：
3.9
通讯作者：
Fu, Jinglin
中科院分区：
文献类型：
--
作者：
Wang, Zhicheng;St. Iago-Mcrae, Ezry;Ebrahimimojarad, Alireza;Won Oh, Sung;Fu, Jinglin
底物限制和沟道在多酶途径中起着关键作用，并被认为影响仿生和人工纳米反应器的催化效率和特异性。在这里，我们报道了一个多酶系统的调节与级联活性的影响，表面亲和力结合底物分子。用适体修饰的DNA折纸被用于在固定化酶的局部区域结合和富集ATP分子，从而将酶级联的活性提高2倍以上。或者，用封闭的适体修饰的DNA纳米结构不与ATP结合，从而降低酶级联的活性。Michaelis-Menten动力学表明，用适体修饰的酶纳米结构的表观Km值降低（降低约3倍），表明由于底物的局部富集，酶附近的有效底物浓度较高。相反，增加的表观Km值（高102倍），观察到的封闭的适体修饰的酶纳米结构，可能是由于排除接近表面的基板。类似的概念，这种改性的表面-基底相互作用应适用于其他多酶系统固定在纳米结构上，这可能是有用的仿生纳米反应器的发展。
Substrate confinement and channeling play a critical role in multienzyme pathways and are considered to impact the catalytic efficiency and specificity of biomimetic and artificial nanoreactors. Here we reported a modulation of a multienzyme system with the cascade activity impacted by the surface affinity binding to substrate molecules. A DNA origami modified with aptamers was used to bind and enrich ATP molecules in the local area of immobilized enzymes, thereby enhancing the activity of an enzyme cascade by more than 2-fold. Alternatively, DNA nanostructure modified with blocked aptamers does not bind with ATP, thereby reducing the activity of the enzyme cascade. The Michaelis–Menten kinetics showed decreased apparentKMvalues (∼3-fold lower) for enzyme nanostructures modified with aptamers, suggesting the higher effective substrate concentration near enzymes due to the local enrichment of substrates. Conversely, increased apparentKMvalues (∼2-fold higher) were observed for enzyme nanostructures modified with blocked aptamers, possibly due to the exclusion of substrates approaching the surface. The similar concept of this modified surface–substrate interaction should be applicable to other multienzyme systems immobilized on nanostructures, which could be useful in the development of biomimetic nanoreactors.